MLKL Double Nickase Plasmid (h): sc-401248-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MLKL Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- MLKL Double Nickase Plasmid (h) and MLKL Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting MLKL. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: MLKL Antibody (3B2): sc-293201
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MLKL Double Nickase Plasmid (h) | sc-401248-NIC | 20 µg | $410.00 | |||
MLKL Double Nickase Plasmid (h2) | sc-401248-NIC-2 | 20 µg | $410.00 |
Mixed lineage kinase domain-like pseudokinase (MLKL) is a terminal effector of necroptosis, activated downstream of RIPK1/RIPK3 signaling to execute regulated membrane permeabilization following inflammatory or death receptor stimuli. Upon phosphorylation by RIPK3, MLKL undergoes oligomerization and translocation to cellular membranes, promoting ion dysregulation and lytic cell death that shapes innate immune responses. MLKL activity intersects with TNF, TLR, and interferon-driven pathways, influencing cytokine release and tissue damage during sterile inflammation and infection. Dysregulated necroptotic signaling involving MLKL has been implicated in models of neuroinflammation, ischemia-reperfusion injury, inflammatory bowel pathology, and tumor-immune interactions, making it a useful node for pathway dissection.
MLKL Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the MLKL locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within MLKL. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt MLKL function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of MLKL-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.